Danielsen E M, Norén O, Sjöström H
Biochem J. 1982 Apr 15;204(1):323-7. doi: 10.1042/bj2040323.
A crude RNA fraction, prepared from pig small intestine, was found to be more efficient than a fraction enriched in polyadenylated RNA in directing translation of polypeptides with Mr greater than 100000 in a rabbit reticulocyte lysate system. Aminopeptidase N (EC 3.4.11.2) synthesized in vitro was immunopurified from the translation mixture and analysed by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis. It was found to have an apparent Mr of 115000 regardless of whether the translation was performed in the absence or presence of proteinase inhibitors. This result contradicts the possibility of aminopeptidase N being synthesized as a large single-chain precursor polypeptide.
从猪小肠制备的粗RNA组分,在兔网织红细胞裂解物系统中指导翻译大于100000的Mr多肽时,被发现比富含聚腺苷酸化RNA的组分更有效。体外合成的氨肽酶N(EC 3.4.11.2)从翻译混合物中免疫纯化,并通过十二烷基硫酸钠/聚丙烯酰胺凝胶电泳进行分析。无论翻译是在不存在还是存在蛋白酶抑制剂的情况下进行,都发现其表观Mr为115000。该结果与氨肽酶N作为大的单链前体多肽合成的可能性相矛盾。